ToRCH IgG/IgM

Assay Development

www.MeridianLifeScience.com
 ToRCH Overview

ToRCH is an acronym for a group of infections that can cause significant birth
defects and even fetal death. Meridian Life Science offers a complete range of
antigens and other reagents for the detection of IgG and IgM antibodies in various
assay formats such as EIA, rapid anti-IgM assays and Immunofluorescence (IFA).
The ToRCH test measures the levels of an infants IgM vs IgG Serology Timeline
antibodies against five groups of chronic infections:
toxoplasmosis, rubella, cytomegalovirus (CMV), herpes
simplex virus (HSV) and other infections. The other
infections usually include syphilis, hepatitis B, coxsackie
virus, Epstein-Barr virus (EBV), varicella-zoster virus
(VZV), and human parvovirus.
These infectious diseases are all associated with
congenital abnormalities resulting from maternal
infection. Although these organisms typically cause only
asymptomatic or mild infection in the mother, they can
have serious consequences for the fetus.
If the infection occurs during the first three months of
pregnancy and if it is a primary infection (newly acquired
during pregnancy), the risk of congenital abnormalities is
much higher as compared to a secondary or reactivated
infection. CMV is the most common cause of For most ToRCH organisms, the initial screening test is
congenital infectious disease with a much higher rate of based on the detection of antibodies to the organism.
transmission (10% vs. 1%) for mothers with a primary Subsequent screening, if required, is carried out using
infection compared to a reactivation. Consequently it is a monoclonal antibody-based immunofluorescent
a very important part of prenatal care to recognize these assay (IFA). Assays are commercially available for the
infections in the first trimester of pregnancy. detection of IgG, IgM, or both IgG/IgM antibodies. In
most cases, IgG reactivity in the absence of IgM
reactivity is indicative of a past infection, while IgM
reactivity in the absence of IgG reactivity indicates a
current infection. However, for some ToRCH diseases
such as toxoplasmosis and CMV infections, IgG avidity
has recently been found to be useful for identifying
primary infections. An IgG antibody produced in the first
few months following an initial infection has a lower
avidity than an IgG antibody produced several months
or years later; consequently, low-avidity antibody can
be used to specifically identify high-risk mothers with
a primary infection. To protect a fetus from ToRCH
infection, early diagnosis through first trimester
screening is critical.

2 | ToRCH REAGENTS - IgG and IgM Assay Development

Meridian ToRCH Antigens & Antibodies
Antigens and Antibodies Assay Reagents

Toxoplasma gondii Anti-human IgM

8122 Native Ag., Grade I with NP40 Z01235M MAb to IgM
EV9519 Native Ag., Grade II W01258G Goat PAb ( chain)
EV8131 Native Ag., Grade III W01259G Goat PAb ( chain), low
R18426 Rec. Ag. p30 (SAG1), E. coli cross-reactivity to IgA & IgG
R01581 Rec. Ag. p35 (GRA8), E. coli
C01523M MAb to T. gondii SAG1 (p30) protein
C01588M MAb to T. gondii (38kDa protein) Anti-human IgG
C01589M MAb to T. gondii (38kDa protein)
Z86238M MAb to IgG (Fc), no cross-reactivity
with IgA, IgM or IgE
Rubella Virus
6075 Native Ag., Grade III, highly pure IgG Absorbent (Required for IgM Assays)
6076/6123 Native Ag., Grade IV, highly pure
L15406G Goat anti-human IgG (Fc)
R01491 Rec. Ag., E1 Mosaic, E. coli
8120 IgM Diluent
EV9525 MAb to Rubella, purified
EV9526 MAb to Rubella, purified
Solid Phase Blocking Buffers
Cytomegalovirus (CMV) J82001B ELISA blocking buffer
J82310B Lateral Flow blocking Buffer, Tris-based
7511 Native Ag., IgM Concentrate
J82300B Lateral Flow blocking Buffer
7600 Native Ag, IgG/IgM Concentrate
J16403D Coating stabilizer and Blocking Buffer
EV9268 Native Ag., Grade II
R01561 Rec. Ag pp52 (UL44), E. coli
R01562 Rec. Ag pp65 (UL83), E. coli
R01563 Rec. Ag pp150 (UL32), E. coli
R01567 Rec. Ag pp38 (UL80a), E. coli
R18103 Rec. Ag pp28 (UL99), E. coli
C86314M MAb to Early Antigen (65kDa protein)
C8A022M MAb to Immediate Early Antigen, pp72

Herpes Simplex Virus (HSV) 1 To order

7305 Native Ag.
Email: orders@meridianlifescience.com
VTI520 Rec. Ag. glycoprotein G 1 (S. cerevisiae)
Phone: +1 901-382-8716
C05014MA MAb to Nucleocapsid protein (155kDa)
Fax: +1 901-333-8223
C66150M MAb to Glycoprotein G 1
Meridian Life Science, Inc.
Herpes Simplex Virus (HSV) 2 5171 Wilfong Rd. / Memphis, TN 38134
7705 Native Ag. www.meridianlifescience.com
VTI530 Rec. Ag. glycoprotein G 2 (S. cerevisiae) ISO 9001:2008 Certified Company
C65019M MAb to Glycoprotein D

www.MeridianLifeScience.com | 3
 Common Types of ToRCH Assays
ToRCH IgG & IgM Capture Assays
A ToRCH serologic test detects IgM and IgG antibodies to the ToRCH panel of infectious diseases (Toxo, Rubella,
CMV and HSV). IgM is the immediate antibody that is produced once a human is exposed to a bacteria, virus
or a toxin and disappears within 2-3 weeks. It is then replaced by IgG which lasts for life and provides lasting
immunity. Meridians ToRCH antigens are suitable for IgG and IgM detection. They can be used in a range of
immunoassay formats including, but not limited to, ELISA, LF, CLIA, rapid assays, and bead-based assays.

Detectable
INDirect General Assay Principle
product
1. Solid-phase (assay plate, beads, etc) is coated with the antigen
Substate 2. Blocking buffer is added to block the remaining binding site
3. Sample is added and patients IgG or IgM antibody binds to
Secondary Enzyme the antigen
Antibodies
4. Detection can either be by direct or indirect methods
Blocking
Patients IgG or IgM Ab Reagent
Antigen
Indirect detection: uses a labeled secondary antibody for
detection. The secondary antibody has specificity for human IgG or IgM.

ToRCH Rapid Anti-IgM Assays

Rapid anti-IgM assays are particularly sensitive in demonstrating IgM responses early in the illness. These assays
work by binding IgM-specific antibodies in the patients specimen to a solid phase coated with an anti-IgM capture
antibody. Soluble antigen is added in excess allowing the specific IgM antibody-antigen reaction to occur in the
absence of competing immunoglobulin isotypes. Finally, a labelled detection antibody is added which has specific
reactivity against the antigen. Assay sensitivity can be highly dependent on the purity of the antigen used.
ELISA capture for IgM determination minimizes interference of rheumatoid factor.

General Assay Principle

Substate
1. Solid-phase is coated with
Detectable
product anti-human IgM (MAb or PAb
Blocking buffer is added to block
Detectable the remaining binding site)
product
Enzyme 2. IgM-specific antibodies in the
patients sample bind to the
anti-human IgM
Anti-ToRCH
3. Antigen (e.g. Rubella, toxo) is
Antigen
added in excess and a antibody-
IgM captured antigen-antibody complex forms
from patient ToRCH Antigen
sample 4. Detection can either be by direct
or indirect methods

Anti-human IgM antibody

Recommended:
Cat# Z01235M
Cat# W01258G
Cat# W01259G
Direct detection INDirect detection
Direct detection: uses a labeled antigen Indirect detection: uses a labeled
that reacts directly with the antibody. secondary antibody for detection.

4 | ToRCH REAGENTS - IgG and IgM Assay Development

How to Increase Assay Sensitivity
Use a solid phase blocking buffer
Solid phase blocking buffers are designed to
Recommended Reagents:
efficiently prevent non-specific binding, reduce
background noise, and stabilize coated proteins
J82100B Blocking Buffer for ELISA
to enable more sensitive immunoassays. They
work by blocking unoccupied spaces on the J82300B Blocking Buffer for Lateral Flow, PBS Based
surface to prevent non-specific binding to this J82310B Blocking Buffer for Lateral Flow, Tris Based
surface by other proteins or biomolecules. J16430D Coating Stabilizer and Blocking Buffer

Use IgG absorbant to remove IgG and RF

The sensitivity and specificity of IgM detection Recommended Reagents:

can be compromised by the presence of IgG
in the patient sample. There are two major
mechanisms by which IgG can interfere with
assays for IgM and cause a false result:
by competing with specific IgM for
substrate binding sites
L15406G Goat anti-human IgG FC (GAHG)
Dilute prior to adding to patient sample.
Recommend diluting 1:10 in PBS.
Add in a ratio of 1:10 to patient sample and allow
to incubate 5-30 minutes.

by forming immune complexes with 8120 IgM Diluent

Rheumatoid Factor (RF) which can compete
with specific IgM for substrate binding sites
Removal of IgG and RF-IgM can be
accomplished by pre-treating the patient
specimen with goat anti-human IgG.
In a separate tube, dilute the patient serum sample
in the IgM Assay diluent at a 1:21 dilution or greater
(mix well). The diluent must be standardized with
the other assay components.

How to use Blocking Buffer and GAH IgG

General Protocol
STEP 1 Optimize the STEP 2 Add blocking buffer STEP 3 Pre-incubate GAHG STEP 4 Add patient sample
plate/nitrocellulose-coating A. Add the blocking solution directly with the patients serum mixture to the reaction well
conditions for the antigen to the wells, blotting membrane A. Dilute GAH IgG 1:10 in PBS A. Incubate patient sample with the
or nitrocellulose membrane B. Add diluted GAH IgG to patient antigen
A. Coat the plate with antigen:
depending on the assay type sample 1:10 and mix B. Perform wash steps: remove
2-10 g/mL solution of protein
being used. C. Incubate for 3-5 min and non-bound reagents
dissolved in an alkaline buffer
Use at 1X concentration or proceed with sample testing
such as phosphate-buffered
with further dilution
saline (pH 7.4) or carbonate-
If patient IgM is
bicarbonate buffer (pH 9.4) ELISA Blocking Buffer:
present it binds
Cat# J82100B to the antigen.
B. Incubate plate for several hours
to overnight at 4-37C Lateral Flow Blocking Buffer:
Cat# J82300B Goat
C. Remove coating solution, Patients
Coating Stabilizer & Blocking anti-human
perform wash steps serum with both IgG (GAHG)
Buffer: Cat# J16430D IgG and IgM
antibodies
B. Incubate at room temperature
Patient sample
for 30 minutes to 2 hours
C. Continue with
your process
Patients serum The GAHG
and reagents and GAHG are complexes
according to the pre-incubated with the
Antigen is assay protocol prior to adding patients STEP 5 Detection by direct
coated to to plate IgG or indirect methods
the plate

www.MeridianLifeScience.com | 5
 Commercial Assays &
Recommended Products
Toxoplasma IgM
Detection Recommended
Assay Type Solid Phase Antigen
System Products
Mouse anti-human IgM, Z01235M,
Abbott CMIA Paramagnetic
F(ab)2 to Toxo & Rec. Direct, CL C01523M
ARCHITECT IgM-capture sandwich method microparticles
p30 antigen and R18426
Z01235M,
Abbott MEIA, Mouse anti-human IgM, MAb
Microparticles Indirect, MUB C01523M
AxSYM IgM-capture sandwich method to Toxo p30 & native lysate
and EV8131
Z01235M,
Siemens CLIA Paramagnetic Mouse anti-human IgM,
Direct, CL C01523M
IMMULITE IgM-capture sandwich method particles MAb to Toxo & p30 antigen
and EV8131
Z01235M,
DiaSorin CLIA Paramagnetic Mouse anti-human IgM, MAb
Indirect, CL C01523M
LIAISON IgM-capture sandwich method particles to Toxo & native antigen
and EV8131
Goat anti-human IgM, MAb W01259G,
BioMrieux ELFA Solid phase
to Toxo p30 & native antigen Direct, MUB C01523M
VIDAS IgM-capture sandwich method receptacles
(RH Sabin strain) and EV8131

Toxoplasma IgG
Detection Recommended
Assay Type Solid Phase Antigen
System Antigens
Abbott Paramagnetic Rec. antigens to 30 and p35 R18426, R01581
CMIA Indirect, CL
ARCHITECT microparticles (GRA8), anti-human IgG and Z86238M
CMIA 2 assays with and without
ARCHITECT Paramagnetic Rec. antigens to 30 and p35 R18426, R01581
blocking buffer to dissociate Indirect, CL
Avidity microparticles (GRA8), anti-human IgG and Z86238M
low-avidity antibodies
Siemens
Paramagnetic
AVIDA CLIA Native p30 antigen Direct, CL EV8131
particles
Centaur
Siemens
Paramagnetic Partially purified antigen,
IMMULITE CLIA Indirect, CL 8122 or EV9519
particles mouse anti-human IgG
2000
DiaSorin Paramagnetic Native antigen, 8122 or EV9519,
CLIA Indirect, CL
LIAISON microparticles anti-human IgG and Z86238M
DiaSorin CLIA 2 assays with and without a
Paramagnetic Native antigen, 8122 or EV9519,
LIAISON 6M urea elution step to dissociate Indirect, CL
microparticles anti-human IgG and Z86238M
Avidity low-avidity antibodies
BioMrieux Solid phase
ELFA Native antigen Direct, MUB 8122 or EV9519
VIDAS receptacles
BioMrieux ELFA 2 assays (reference and test)
Solid phase Native antigen (RH Sabin 8122 or EV9519
VIDAS with and without a disassociation Direct, CL
receptacles strain), anti-human IgG and Z86238M
Avidity wash step to remove low-avidity

ELFA: Enzyme-linked fluorescence assay

CLIA: Chemiluminescent Immunoassay
CMIA: Chemiluminescent Microparticle Immunoassay
MUB: Methyl-umbellifery
CL: Chemiluminescence
MEIA: Microparticle Enzyme Immunoassay
ECLIA: Electrochemiluminescence Immunoassay

6 | ToRCH REAGENTS - IgG and IgM Assay Development

Rubella IgM
Detection Recommended
Assay Type Solid Phase Antigen
System Products
Abbott
Paramagnetic Viral lysate (strain HPV77) and
ARCHITECT CMIA Indirect, CL 6076/6123
microparticles mouse anti-human IgM MAb
i2000SR
Recombinant rubella like
Roche R01491 and
CLIA IgM-capture sandwich method Magnetic beads particles and rubella-specific Indirect, CL
ELECSYS EV9525 or EV9526
antibodies
Siemens
Polystyrene
IMMULITE CLIA IgM-capture sandwich method Viral lysate (strain HPV77) Indirect, CL 6076/6123
beads
2000
Siemens
Paramagnetic Viral lysate and rubella 6076/6123 and
AVIDA CLIA IgM-capture sandwich method Indirect, CL
particles specific antibody EV9525 or EV9526
Centaur XP

Rubella IgG
Detection Recommended
Assay Type Solid Phase Antigen
System Products
Abbott Partially purified viral lysate
Paramagnetic
ARCHITECT CMIA (strain HPV77), mouse Indirect, CL 6075 and Z86238M
microparticles
i200SR anti-human IgG MAb
Abbott Purified viral lysate and
MEIA Microparticles Indirect, MUB 6075 and Z86238M
AxSYM anti-human IgG
Siemens Purified viral lysate (strain
Paramagnetic
AVIDA CLIA moderate complexity HPV77) and mouse anti- Indirect, CL 6075 and Z86238M
particles
Centaur XP human IgG FC
Siemens
Paramagnetic
IMMULITE CLIA moderate complexity Viral lysate Indirect, CL 6075
particles
2000
Beckman Paramagnetic
EIA Viral lysate Indirect, CL 6075
ACCESS 2 particles
Recombinant E1 antigen,
Roche CLIA, double antigen sandwich with R01491 and
Magnetic beads rubella like particles and Indirect, CL
ELECSYS an IgG-capture method EV9525 or EV9526
rubella specific antibodies
BioMrieux ELFA, sandwich immunoassay Solid phase Viral lysate (strain MR383)
Indirect, MUB 6075 and Z86238M
VIDAS method receptacles and anti-human IgG
Ortho Viral lysate and mouse-anti- Indirect,
CLIA high complexity Plastic wells 6075 and Z86238M
VITROS Eci human IgG Luminescence

CMV IgM
Detection Recommended
Assay Type Solid Phase Antigen
System Products
Viral lysate (strain AD169),
Abbott Paramagnetic R01561, R01563
CMIA rec. antigen pp150 and pp52, Indirect, CL
ARCHITECT microparticles and W01259G
and anti-human IgM
Rec. antigen CMV pp150, R01561, R01562,
Abbott
MEIA Microparticles pp52, pp65, pp38, and Indirect, MUB R01563, R01567
AxSYM
anti-human IgM and W01259G
Siemens Purified antigen (strain 7600 or 7511 and
CLIA 3 step assay, IgM-capture Paramagnetic
IMMULITE AD169), Goat anti-human IgG, Direct, CL W01259G and
sandwich method particles
2000 goat anti-human IgM L15406G
Roche ECLIA IgM-capture sandwich Paramagnetic Rec. antigens CMV pp150 and R01561, R01563
Direct, CL
ELECSYS method microparticles pp52, and anti-human IgM and Z01235M
BioMrieux Solid phase Viral lysate (strain AD169), 7600 or 7511 and
ELFA IgM-capture sandwich method Indirect, MUB
VIDAS receptacles mouse anti-human IgM Z01235M

www.MeridianLifeScience.com | 7
 Commercial Assays &
Recommended Products continued
CMV IgG
Detection Recommended
Assay Type Solid Phase Antigen
System Antigens
Abbott
Paramagnetic
ARCHITECT CMIA Viral lysate Indirect, CL 7600 or 7511
microparticles
i2000SR
CMIA, two assays with and
Abbott
without liquid CMV antigen to Paramagnetic 7600 or 7511 and
ARCHITECT Viral lysate, anti-human IgG Indirect, CL
neutralize high-avidity CMV microparticles Z86238M
Avidity
antibodies
Abbott AxSYM MEIA Microparticles Viral lysate Indirect, MUB 7600 or 7511
Siemens
Paramagnetic
IMMULITE CLIA Purified Antigen Indirect, CL 7600 or 7511
particles
2000
R01561, R01563,
Roche ECLIA, one step double sandwich Paramagnetic Rec. antigens CMV pp150,
Direct, CL R01567 and/or
ELECSYS method microparticles pp52, pp28, pp38
R18103
Roche ECLIA, two assays with and R01561, R01563,
Paramagnetic Rec. antigens CMV pp150,
ELECSYS without chaotropic conditions to Direct, CL R01567 and/or
microparticles pp52, pp28, pp38
Avidity dissociate low-avidity antibodies R18103
BioMrieux Solid phase 7600 or 7511 and
ELFA two step sandwich method Viral lysate, anti-human IgG Direct, MUB
VIDAS receptacles Z86238M
ELFA two assays with and without
BioMrieux Solid phase 7600 or 7511 and
6 M urea to dissociate low-avidity Viral lysate, anti-human IgG Direct, MUB
VIDAS Avidity receptacles Z86238M
antibodies

HSV-1 & HSV-2 IgG/IgM

Detection Recommended
Assay Type Solid Phase Antigen
System Antigens
DiaSorin
LIAISON Magnetic Rec. HSV-1 gG 1 and
CLIA, two steps Indirect, CL VTI520, Z86238M
HSV-1 Type particles mouse anti-human IgG
Specific
DiaSorin
LIAISON Magnetic Rec. HSV-2 gG 1 and mouse
CLIA, two steps Indirect, CL VTI530, Z86238M
HSV-2 Type particles anti-human IgG
Specific
Focus PLEXUS
Rec. HSV-1 gG 1, HSV-2 gG 1 Indirect, VTI520, VTI530
HerpesSelect EIA multiplex flow three steps Beads
and goat anti-human IgG Fluorescent and L15406G
1 and 2 IgG
Rec. gG1 and synthetic
Biorad
peptide p8C:BSA derived from
BIOPLEX 2200 Carboxy-coated Direct, VTI520, VTI530,
EIA multiplex flow three steps gG2 sequence (patented) and
HSV-1 & dyed beads Fluorescent and Z86238M
mouse anti-human IgG and
HSV-2 IgG
mouse anti-human FXIII
Roche
ECLIA, two step Paramagnetic
ELECSYS Rec. HSV-1, HSV-1 viral lysate Direct, CL VTI520 and 7305
antigen sandwich microparticles
HSV-1 IgG
Siemens
HSV-1 (strain MacIntyre),
Immunlite Paramagnetic 7305, 7705
CLIA HSV-2 (strain G) viral lysate Indirect, CL
2000 Herpes particles and Z86238M
and anti-human IgG
1/2 IgG

Meridian Life Science, Inc.

5171 Wilfong Road | Memphis, TN 38134
8 901.382.8716 800.327.6299 April 2015